The Mcm2-7 complex has in vitro helicase activity.

Abstract:

:Helicases unwind duplex DNA ahead of the polymerases at the replication fork. However, the identity of the eukaryotic replicative helicase has been controversial; in vivo studies implicate the ring-shaped heterohexameric Mcm2-7 complex, although only a specific subset of Mcm subunits (Mcm467) unwind DNA in vitro. To address this discrepancy, we have compared both Mcm assemblies and find that they differ in their linear single-stranded DNA association rate and their ability to bind circular single-stranded DNA. These differences depend upon the Mcm2/5 interface, which we hypothesize serves as an ATP-dependent "gate" within Mcm2-7. Importantly, we find that reaction conditions that putatively close the Mcm2-7 "gate" reconstitute Mcm2-7 helicase activity. Unlike Mcm467, Mcm2-7 helicase activity is strongly anion dependent. Our results show that purified Mcm2-7 acts as a helicase, provides functional evidence of a Mcm2/5 gate, and lays the foundation for future mechanistic studies of this critical factor.

journal_name

Mol Cell

journal_title

Molecular cell

authors

Bochman ML,Schwacha A

doi

10.1016/j.molcel.2008.05.020

subject

Has Abstract

pub_date

2008-07-25 00:00:00

pages

287-93

issue

2

eissn

1097-2765

issn

1097-4164

pii

S1097-2765(08)00398-5

journal_volume

31

pub_type

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